An HPLC method has been optimized and validated for determining swainsonine, a potent glucosidase inhibitor present in locoweeds (any of several plants of the genera Oxytropis and Astragalus in the pea family, which can cause severe poisoning when eaten by livecstock.) of China with isopropanol as a weak eluent.Swainsonine was separated from an extract of locoweeds on hydrophilic interaction chromatography column, using isopropanol-2 mM ammonium acetate (52:48, v/v) as mobile phase at a flow rate of 0.4 mL/min.During post column detection, swainsonine was detected by evaporative light scattering detector.The method was validated and the detection limit was 6 µg/mL.This method is sufficiently sensitive for determining swainsonine in Chinese locoweeds.
The investigation of the effect of Fructus arctii extract on anti-tumor of mice bearing uterine cervical carcinoma(U_(14))was studied.The effect of Fructus arctii extract on the inhibition rate of solid tumor and the life span of ascites tumor were conducted with the experiment model in vivo of animal tumor.Compared with the control group,the Fructus arctii extract(1?000,2?000 and 4?000 mg/kg body weight) and CTX(25 mg/kg body weight) significantly reduced the tumor weight of solid tumor and increased the life span of ascites tumor harboring mice(P0.01).The inhibition rate of solid tumor and the rate of life span were up to 31.7, 49.7,57.4 and 83.6%,and 39.8,46.6,62.5 and 91.6%,respectively.The result indicated that Fructus arctii extract had antitumor action.
【Objective】The aim of our research was to investigate the effect of Patrinia heterophylla Bunge polysaccharide(PHB-P)on U14-bearing mice.【Method】The tumor inhibition ratio was calculated by the model U14-bearing tumor mice.Subcutaneous injection of normal saline and given with cyclophosphamide 25mg/kg were used as negative and positive controls,treated with PHB-P(30,60,90mg/kg)for 14 days.Detection of apoptosis was performed by TUNEL staining assays.The activity of serum LDH and AKP was examined using the kits.Imunohistochemical analysis assay was applied on the protein expression productions of apoptosis-associated mutant p53,Bcl-2and Bax genes in tumor tissues.【Result】The results showed that the groups of PHB-P(60,90mg/kg),compared with the negative control group,significantly decreased tumor weight and the difference was significant(P0.05);compared with the negative control group,the number of apoptotic cells of the PHB-P(60,90mg/kg)groups significantly increased(P 0.05);compared with the negative control group,tumor bearing mice serum LDH activity of the groups of PHB-P(60,90mg/kg)significantly decreased(P0.05);In each group of PHB-P(30,60,90mg/kg),ser-um AKP activity increased compared with the negative control group,but the difference was not significant(P0.05);Bax protein expression was significantly higher than that of the negative control group(P 0.05),mutant p53and Bcl-2protein expression in PHB-P(60,90mg/kg)group compared with the negative control group decreased significantly(P0.05).Compared with the positive control group,the effects of anti-tumor activity of the PHB-P(60,90mg/kg)was close to the clinical chemotherapeutic agent cyclophosphamide.【Conclusion】Our data suggested that PHB-P might induce tumor cells apoptosis and inhibit tumor growth,including regulation of the apoptosis-associated genes.
YLZZ-2 is a strain of Stenotrophomonas maltophilia capable of Swainsonine(SW) degradation.In this study,four condition factors were optimized by orthogonal test including initial culture medium pH,temperature,SW substrate concentration,and speed of shaking.The results show that YLZZ-2 could degrade SW at its best efficiency under the condition of pH value 7.0,temperature 30℃,SW concentration 5%,and shaking speed 160 r·min-1.Results of resistance test show that the bacteria had ampicillin-and-kanamycin resistance.The resistance disappeared and the ability of SW degradation lost when the plasmids were eliminated from YLZZ-2,which indicates that the SW degradation function of YLZZ-2 is associated with its plasmids.
Arthrobacter sp. HW08 is capable of swainsonine (SW) degradation with high efficiency. The aim of this study was to screen the degradative plasmids from Arthrobacter sp. HW08 and investigate their ability of degrading SW. A genomic library of Arthrobacter sp. HW08 was successfully constructed by BamHI restriction digestion of genomic DNA and pUC19 vector, ligation and transformation into DH5. Five plasmids of different sizes were screened from 100 genomic library strains, and were named as pUCSW-1, pUCSW-2, pUCSW-3, pUCSW-4 and pUCSW-5, and their insertion elements had sizes of 6 684, 5 093, 136, 1 452 and 2 082 bp, respectively. They contained 7 major open reading frames. SW (400 mg l -1 ), as only carbon source, cultivated with the mixture of the five plasmid-transformants was degraded within 6 h. The degrading capability was equivalent to that of strain HW08. SW degradative ability of minimum combinations was pUCSW-2, 3, 5 > pUCSW-1, 2, 4 > pUCSW-1, 2, 5 > pUCSW-1, 3, 5, which were all slower than that of strain HW08. Any two or single transformant had no degradative ability. The results of this study support the search of the key genes of SW degradation and the investigating mechanism of SW biodegradation or exploration of its metabolic pathway, and even construction of transgene engineering bacteria to protect animals from lesion when locoweed is consumed. Key words : Swainsonine, biodegradation, degradative plasmids, Arthrobacter sp. HW08.
The research investigated the effect of Patrinia heterophylla Bunge (Valerianaceae) polysaccharides (PHB-P1) on U14-bearing mice. The tumor weight of mice treated with PHB-P1 (30, 60 mg/kg body weight) was significantly lower than that of the control group, a decrease of serum lactate dehydrogenase (LDH) activity was observed, and the serum alkaline phosphatase (AKP) level was increased slightly. The number of apoptotic tumor cells was significantly increased in the mice by treatment of PHB-P1 (30, 60 mg/kgbw). Cell cycle analysis showed the accumulation of tumor cells in the G2/M phase and a relative decrease of the S phase. By the immunohistochemical analysis, PHB-P1 (30, 60 mg/kgbw) might up-regulate the expression of p53 and Bax, and significantly inhibited the expression of Bcl-2 in tumor tissues. In conclusion, PHB-P1 could inhibit tumor growth and induce tumor cell apoptosis.
【Objective】 The study established a method for determination of Swainsonine.【Method】 Swainsonine in Astragalus locoweed was determinated by mainternal standard gas chromatography with me-Gal as internal standard under the oven temperature 200 ℃,injection temperature 300 ℃,detector(FID) temperature 280 ℃ and flow rate of carrier gas 200 kPa.【Result】 The analysis results of single-calibration-ratio was 1.067 8 and good linear relationship was obtained in the range of 0.047 to 6 mg/mL.The recovery percentage was 107.33%.The SW contents of A.variabilis and A.strictus were (39.2±0.16) mg/kg and(43.6±0.13) mg/kg respectively.【Conclusion】 The internal standard method had high accuracy and good repertability and could be a determination method of Swainsinine.
OBJECTIVE To study the antitumor effect of the stings of Gleditsia sinensis on mice bearing uterine cervical carcinoma (U14) and the expression of PCNA (proliferating cell nuclear antigen) and p53. METHOD The effect of the ethanolic extract of G. sinensis stings on the inhibition rate of solid tumor and the life span of ascites tumor were calculated by the animal tumor model experiment in vivo. The positive cell numbers of PCNA and mutant p53 protein were measured by immunohistochemical SP method. RESULT As compared with the control group, the ethanolic extract of G. sinensis stings (250, 500 and 1 000 mg x kg(-1) body weight, p.o.) and CTX (25 mg kg(-1) body weight, i.p.) administration significantly reduced the tumor weight of solid tumor and increased the life span of ascites tumor harboring mice (P < 0.01). The inhibition rate of solid tumor and the rate in life span were up to 47.44%, 59.49%, 63.92%, 73.42% and 52.21%, 67.26%, 78.76%, 95.58% respectively. Meanwhile,the expression of PCNA and mutant p53 protein also suppressed by ethanolic extract (P < 0.05, P < 0.01). CONCLUSION The stings of G. sinensis showed antitumor activity and its possible mechanism might be related with the expression inhibition of PCNA and mutant p53 protein.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)
 scale(5.56106)'%3e%3ccircle r='1'/%3e%3cg id='d'%3e%3cg id='c'%3e%3cpath id='b' d='M.195-.98 0-1.39l-.195.408' transform='rotate(11.25)'/%3e%3cuse xlink:href='%23b' transform='rotate(22.5)'/%3e%3cuse xlink:href='%23b' transform='rotate(45)'/%3e%3c/g%3e%3cuse xlink:href='%23c' transform='rotate(67.5)'/%3e%3c/g%3e%3cuse xlink:href='%23d' transform='scale(-1 1)'/%3e%3c/g%3e%3cg transform='translate(0 58.787) scale(8.1434)'%3e%3ccircle r='1'/%3e%3cg id='g'%3e%3cg id='f'%3e%3cpath id='e' d='M.259.966 0 1.576l-.259-.61'/%3e%3cuse xlink:href='%23e' transform='rotate(180)'/%3e%3c/g%3e%3cuse xlink:href='%23f' transform='rotate(90)'/%3e%3c/g%3e%3cuse xlink:href='%23g' transform='rotate(30)'/%3e%3cuse xlink:href='%23g' transform='rotate(60)'/%3e%3c/g%3e%3c/g%3e%3c/svg%3e)